The effect of covalent cross-links between the membrane components of microcapsules on the dissemination of encapsulated malignant cells.

Stem cells and immortalized cells have considerable therapeutic potential but present risks of malignant transformation. Cell microencapsulation allows transplantation without immunosuppression. We have developed a method for microencapsulating living cells within covalently cross-linked membranes that are chemically and mechanically extremely resistant. We provide herein direct evidence that these microcapsules can prevent malignant cell dissemination. When 20,000 or more nonencapsulated EL-4 thymoma cells were implanted intraperitoneally in mice, all recipients died with widespread metastasis within 26.3+/-1.0 days. All recipients of 250,000 EL-4 cells microencapsulated in covalently cross-linked membranes were living and disease-free, 150 days post-implantation. Encapsulation in standard microcapsules only slightly delayed the recipient death. Pancreatic islets transplanted using either type of microcapsule presented similar survival. We conclude that microencapsulation in covalently cross-linked membranes prevents malignant cell dissemination.

[1]  R. C. Johnson,et al.  In vivo delivery of recombinant human growth hormone from genetically engineered human fibroblasts implanted within Baxter immunoisolation devices , 1999, Journal of Molecular Medicine.

[2]  P. Chang,et al.  Gene Therapy for Hemophilia , 2000, Artificial cells, blood substitutes, and immobilization biotechnology.

[3]  Li Li,et al.  Bone marrow–derived stem cells initiate pancreatic regeneration , 2003, Nature Biotechnology.

[4]  Wim E. Hennink,et al.  Novel crosslinking methods to design hydrogels , 2002 .

[5]  Y. Lepage,et al.  Studies on Small (<300 μm) Microcapsules: II — Parameters Governing the Production of Alginate Beads by High Voltage Electrostatic Pulses , 1994, Cell transplantation.

[6]  P. Chang,et al.  Combined immunotherapy and antiangiogenic therapy of cancer with microencapsulated cells. , 2004, Human gene therapy.

[7]  P. de Vos,et al.  MTS colorimetric assay in combination with a live-dead assay for testing encapsulated L929 fibroblasts in alginate poly-L-lysine microcapsules in vitro. , 2002, Artificial organs.

[8]  A. Monaco,et al.  An improved method for isolation of mouse pancreatic islets. , 1985, Transplantation.

[9]  D. Kooy,et al.  Clonal identification of multipotent precursors from adult mouse pancreas that generate neural and pancreatic lineages , 2004, Nature Biotechnology.

[10]  P. Bruheim,et al.  Alginate polycation microcapsules. I. Interaction between alginate and polycation. , 1996, Biomaterials.

[11]  Michael L. Nicholson,et al.  Pancreatic Islet Cell Transplantation , 2003 .

[12]  J. Itskovitz‐Eldor,et al.  Insulin production by human embryonic stem cells. , 2001, Diabetes.

[13]  R. Robitaille,et al.  Microencapsulation of living cells in semi-permeable membranes with covalently cross-linked layers. , 2005, Biomaterials.

[14]  G Orive,et al.  Long-term expression of erythropoietin from myoblasts immobilized in biocompatible and neovascularized microcapsules. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[15]  C. Maltin,et al.  Secretion of bioactive human insulin following plasmid-mediated gene transfer to non-neuroendocrine cell lines, primary cultures and rat skeletal muscle in vivo. , 2002, The Journal of endocrinology.

[16]  R. Morgan,et al.  Ex vivo fibroblast transduction in rabbits results in long-term (>600 days) factor IX expression in a small percentage of animals. , 1998, Human gene therapy.

[17]  H. Mashima,et al.  Formation of insulin-producing cells from pancreatic acinar AR42J cells by hepatocyte growth factor. , 1996, Endocrinology.

[18]  S. Efrat,et al.  Murine Insulinoma Cell Line With Normal Glucose-Regulated Insulin Secretion , 1993, Diabetes.

[19]  G. Korbutt,et al.  Stem cells: a promising source of pancreatic islets for transplantation in type 1 diabetes. , 2003, Current topics in developmental biology.

[20]  Giovanni Luca,et al.  Microencapsulated pancreatic islet allografts into nonimmunosuppressed patients with type 1 diabetes: first two cases. , 2006, Diabetes care.

[21]  Peter J. Donovan,et al.  The end of the beginning for pluripotent stem cells , 2001, Nature.

[22]  G. Korbutt,et al.  Insulin expressing cells from differentiated embryonic stem cells are not beta cells , 2004, Diabetologia.

[23]  F. Lim,et al.  Microencapsulated islets as bioartificial endocrine pancreas. , 1980, Science.

[24]  R. Bottino,et al.  Gene- and cell-based therapeutics for type I diabetes mellitus , 2003, Gene Therapy.

[25]  J. Reig,et al.  Insulin-secreting cells derived from embryonic stem cells normalize glycemia in streptozotocin-induced diabetic mice. , 2000, Diabetes.

[26]  G. Wolters,et al.  Effect of Alginate-Polylysine-Alginate Microencapsulation on In Vitro Insulin Release From Rat Pancreatic Islets , 1991, Diabetes.

[27]  Seung K. Kim,et al.  Growth inhibitors promote differentiation of insulin-producing tissue from embryonic stem cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[28]  R. van Schilfgaarde,et al.  Causes of limited survival of microencapsulated pancreatic islet grafts. , 2004, The Journal of surgical research.

[29]  Takahisa Fujikawa,et al.  Teratoma formation leads to failure of treatment for type I diabetes using embryonic stem cell-derived insulin-producing cells. , 2005, The American journal of pathology.

[30]  G. Mattson,et al.  A practical approach to crosslinking , 1993, Molecular Biology Reports.

[31]  L. Rosenberg,et al.  Identification and characterization of small cells in the adult pancreas: potential progenitor cells? , 2002, Cell and Tissue Research.

[32]  F. Pattou,et al.  Expression of progenitor cell markers during expansion of sorted human pancreatic beta cells. , 2005, Gene expression.

[33]  I. Kojima,et al.  Regenerative medicine of the pancreatic β cells , 2005 .

[34]  P. Chang Encapsulation for Somatic Gene Therapy , 1999, Annals of the New York Academy of Sciences.

[35]  R. Robitaille,et al.  Insulin-like growth factor II allows prolonged blood glucose normalization with a reduced islet cell mass transplantation. , 2003, Endocrinology.

[36]  N. Déglon,et al.  Survival of encapsulated human primary fibroblasts and erythropoietin expression under xenogeneic conditions. , 2004, Human gene therapy.

[37]  R. C. Johnson,et al.  Correction of diabetic nod mice with insulinomas implanted within Baxter immunoisolation devices , 1999, Journal of Molecular Medicine.